in-vivo real time non-invassive imaging for longitudinal ... · in-vivo real time non-invassive...
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In-vivo real time non-invassive imaging for longitudinal translational research
Milan Kopecek
Regional Sales Manager
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• Micro-ultrasound
• What is Micro-ultrasound?
• Visualization, measurement and analysis with the Vevo 2100
• Photoacoustics
• What is Photoacoustics?
• Visualization, measurement and analysis with the Vevo LAZR
• Applications
Agenda
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Total Small Animal Solution
• Real-time
• Longitudinal Studies
• Measure Physiological parameters
• Contrast/Molecular Imaging
• Translatable to man
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How Does Micro-Ultrasound Work?
How Does Micro-Ultrasound Work?
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Micro-ultrasound (mouse fetus)
High-frequency = High-resolution
3 - 15 MHz
20cm
30 – 80 MHz
3cm
Melon
Coffee Bean
Conventional clinical ultrasound (human fetus)
Micro-Ultrasound: Clarity
200 – 300 micron resolution
30 micron resolution
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Anatomy and Morphology
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Cardiac Imaging: EKV
• High Temporal Resolution Left Ventricle in Adult Mouse at
10,000 frames per second
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Visualization Measurement Analysis
• Tissue/organ/tumor identification
• Vasculature density, structure and flow
• Biomarkers
• Linear, area and volume measurements
• Blood flow velocity and vessel density
• Cardiac measurements
• Contrast Quantification including biomarker expression
• Heart wall motion and strain
Imaging with the Vevo
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Visualization with Micro-Ultrasound
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1 mm *WAP-TAg Mouse Model
Image courtesy of Cotarla I. and Furth, P., Lombardi Cancer Centre, Georgetown University
Early detection and Quantification of Pre-Palpable Mammary Tumors
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Image courtesy of T. Minko, Rutgers, The State University of New Jersey
Video Loop
3D Quantification of Human Ovarian Tumor Model
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Power Doppler Mode Imaging
Day 1 Day 4
PV = 3.18% PV = 11.22%
Images courtesy of Hastie, Chambers, Lacefield and Fenster; Robarts Research Institute, London
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Precise interventions without the need for invasive surgery
Ultrasound Image-Guided Injections
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Measurement with Micro-Ultrasound
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Quantification of size, vasculature and perfusion
Early detection/screening
Perfusion Volumetrics
Vascularity
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Cardiovascular measurements
LV analysis
Vascular stiffness
Blood flow
Strain
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Analysis with Micro-Ultrasound
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User defined regions of interest
Curve fit algorithms
Vevo MicroMarker Quantification
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Velo
city D
isplacem
ent
Strain
S
train R
ate
Kovacs, Washington University
VevoStrainTM
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• Micro-ultrasound
• What is Micro-ultrasound?
• Visualization, measurement and analysis with the Vevo 2100
• Photoacoustics
• What is Photoacoustics?
• Visualization, measurement and analysis with the Vevo LAZR
• Applications
Agenda
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The Photoacoustic Effect
1) Nanosecond laser pulse illumination
3) Emitted pressure (sound) wave
4) Detection of ultrasound and creation of image
2) Optical absorption, heating and thermoelastic expansion
Optical Ultrasound
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The Photoacoustic Effect
Light in
Sound out
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5 mm 5 mm
5 mm
Meaningful images with the sensitivity of optical imaging and the resolution of ultrasound
The Photoacoustic Imaging
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Visualization Measurement Analysis
• Blood signal
• Vascular density, structure
• Angiogenesis
• Oxygen saturation and total hemoglobin
• Photoacoustic signal intensity (contrast agent measurement)
• Absorption spectra
• Spectral unmixing
Imaging with the Vevo LAZR Photoacoustic System
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Visualization with Photoacoustics
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• Inherent co-registration of photoacoustic and anatomical images – Simultaneous registration of photoacoustic image on 2D and 3D planes – Real-time acquisitions for true in vivo monitoring
What is this signal?
Where is this signal?
Needle Skin Tumor Needle Skin Tumor
Delivery of nanoparticles into tumor
The Photoacoustic Imaging
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Tumor vasculature imaging
Single wavelength imaging of endogenous signal
•Sensitive detection of blood signal
•Functional and non-functional vessels, blood pools
•2D and 3D
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Measurement with Photoacoustics
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( HbT = HbO2 + Hb )
Total Hemoglobin = Oxygenated Hemoglobin + Deoxygenated Hemoglobin
Near-Infrared (NIR)
Oxygen Saturation Calculation
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0
10
20
30
40
50
60
70
80
90
0 100 200
Oxyg
en
Satu
rati
on
(%
)
Time (s)
Region 1 Region 2
100% O2 100% O2 5% O2
Change in Oxygen Saturation in Tumor
Region 1
Region 2
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Analysis with Photoacoustics
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Endogenous contrast
Contrast
Agent
Applications
Vevo LAZR Contrast Applications
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Absorption Spectra Analysis – Spectro Imaging
Indocyanine green absorption spectra measured by photoacoustic imaging from 680 to 970 nm wavelengths.
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Applications of Ultrasound and Photoacoustics
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Single Walled Carbon Nanotubes conjugated to RGD peptide target αvβ3 integrins
(Gambhir lab, Stanford University)
(Heinmiller, et al. EMIM 2011)
Targeted carbon nanotubes
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Change in Measured Photoacoustic Signal over time after carbon nanotube injection
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Normal
Color Doppler Oxygen
saturation Total
hemoglobin
Stroke 2
Stroke 1
Stroke Imaging – Skin removed
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Melanin
Oxygenated blood
3D Imaging of Human Finger
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Pharmacokinetics with Photoacoustics
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Identifying IR800-2DG in the kidney
Peak ~785nm Peak 760nm
Control and kidney
regions have peak PA
signal at different
wavelenths
Peak ~785nm
IR800-2DG in phantom
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Spectral unmixing
Pre
24hrs post
6hrs post
Blue = DeoxyHb Red = OxyHb Green = IR800 2_DG
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Quantifying IR800-2DG in the kidney
0
0.2
0.4
0.6
0.8
1
1.2
1.4
Pre 6 hrs post 24 hrs post
3D quantification of PA signal in the kidney pre, 6hrs and 24hrs post-IR800-
2DG
780nm 850nm
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Methylene blue i.v. Injection – kidney imaging • Subtraction image of bolus injection of methylene blue using the ‘invisible
table’ and trifurcated fiber
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Methylene blue i.v. Injection – kidney imaging
Pre-bolus
7 mins Post-bolus
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Methylene blue i.v. Injection – kidney imaging
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Reporter gene imaging with Photoacoustics
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Photoacoustic Imaging of an Inducible Reporter
• Tyrosinase, the enzyme responsible for melanin production was used as an inducible reporter
• Animals with TYR- and TYR+ tumors on each flank were imaged before and after induction of tyrosinase expression
Abs.
Figure modified from Murkin and Arango. Near-infrared spectroscopy
as an index of brain and tissue Oxygenation. Br J Anaesth 2009; 103 (Suppl. 1): i3–i13
-TYR
+TYR
Absorption spectra for hemoglobin and
melanin
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Post-induction Pre-induction
TYR-
a)
Photoacoustic Imaging of an Inducible Reporter
TYR+
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a)
Photoacoustic Imaging of an Inducible Reporter
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a)
Multi-modality Molecular Imaging with Tyrosinase as a Reporter Gene
MRI PET
PA
Figures from Cheng, Z. et al. Tyrosinase as a multifunctional reporter gene for Photoacoustic/MRI/PET triple modality molecular imaging. Sci Rep. 2013 Mar 19;3:1490
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Imaging Enzymatic Activity
Pre
Post
680 nm 750 nm
Min
Max Pre Injection
680-750
Post Injection
680-750
Subtraction of Images
Levi, J. et al. Clin Cancer Res, 2013
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MSCs + NTs Control MSCs + NTs Control
MSCs + NTs
in vivo monitoring MSC’s labeled with nanotracers
Nam et al SPIE 2012
Ischemic muscle
PEGylated fibrin gel
MSCs labeled with
NTs
US/PA imaging
PEGylated fibrin gel with MSCs +
NTs (105 cells/mL)
MSCs+NTs
Skin
HbO2
Hb
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Longitudinal in vivo monitoring MSC’s labeled with nanotracers
Nam et al SPIE 2012
PEGylated fibrin gel with MSCs +
NTs
DAY 0 DAY 3 DAY 10 DAY 7
Injection
3D combined ultrasound and spectroscopic images of the gastrocnemius in which the
PEGylated fibrin gel containing MSCs/NTs was injected (Day 3,7, and 10)
23 mm x 12.5 mm x 25 mm
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Neuroimaging with Photoacoustics
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Brain imaging with photoacoustics
• Mouse brain coronal section
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Non-Invasive 2D Brain Imaging with Oxyhemo Mode – Prototype LZ201
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PCA
PCommA.
ICA
ICA
Noninvasive imaging of mouse brain
From: Dorr, et al. Neuroimage, 2007
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Non-Invasive 3D Brain Imaging with Oxyhemo Mode
Superior sagittal sinus
Inferior cerebral
vein
Skin intact Skin intact
OxyZated Oxygen saturation map
HemoMeaZure Total hemoglobin map
1mm 1mm
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Imaging of mouse brain ventricles
LZ250 LZ550
Methylene Blue
Injection site
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Imaging of mouse brain ventricles
MRI microscope Multiplexed Vevo LAZR
PA image showing methylene blue in the ventricles
Van Hagen et al., Neurobiology of Disease, 2007.
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Brain Tumor Imaging with Microultrasound and
Photoacoustics
Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility
Scott Floyd lab
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Nonlienar contrast imaging of perfusion with microbubbles
Nonlinear contrast Mouse #1
Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT
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Nonlienar contrast imaging of perfusion with microbubbles
Nonlinear contrast
Hypoperfused area
Mouse #1
Contrast quantifiation (relative blood volume)
Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT
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Multi-modality imaging of hemodynamics in the brain
Potential hypoxic area Mouse #1
Perfusion Flow Oxygenation Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT
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Multi-modality imaging of hemodynamics in the brain
Mouse #1
Perfusion (Relative blood volume, flow estimates)
Flow (Percent vascularity)
Oxygenation (sO2, HbT)
Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT
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Mouse #1
0
10
20
30
40
50
60
70
80
90
100
10 20 30 40 50 60
sO2
Avg
(%
)
Coronal slice # (caudal to rostral)
Avg sO2 in L and R Hemisphere (mouse 1) Caudal to rostral slices
L Hemisphere sO2
R Hemisphere sO2
Quantification of oxygen saturation in L vs R hemispheres
Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT
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Nonlinear contrast Mouse #2
Nonlienar contrast imaging of perfusion with microbubbles
Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT
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Nonlinear contrast
Hypoperfused area
Mouse #2
Hyperperfused area
Hypoperfused area
Contrast quantifiation (relative blood volume)
Nonlienar contrast imaging of perfusion with microbubbles
Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT
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Mouse #2 Potential hypoxic area
Perfusion Flow Oxygenation
Multi-modality imaging of hemodynamics in the brain
Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT
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Mouse #2
Perfusion (Relative blood volume, flow estimates)
Flow (Percent vascularity)
Oxygenation (sO2, HbT)
Multi-modality imaging of hemodynamics in the brain
Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT
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Mouse #2
0
10
20
30
40
50
60
70
80
90
100
10 20 30 40 50
sO2
Avg
(%
)
Coronal slice # (caudal to rostral)
Avg sO2 in L and R Hemisphere (mouse 2) Caudal to rostral slices
L Hemisphere sO2
R Hemisphere sO2
Quantification of oxygen saturation in L vs R hemispheres
Images courtesy of: Swanson Biotechnology Center Animal Imaging and Preclinical Testing Facility and laboratory of Scott Floyd, Koch Institute for Integrative Cancer Research, MIT
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Stroke imaging with Microultrasound and
Photoacoustics
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Stroke vs normal animal - quantification
0
10
20
30
40
50
60
70
80
90
Stroke 1 Stroke 2 Normal
sO
2 (
%)
3D oxygen saturation
0
5000
10000
15000
20000
25000
Stroke 1 Stroke 2 Normal
Hb
T (
a.u
.)
3D total hemoglobin
0%
2%
4%
6%
8%
10%
12%
14%
16%
Stroke 1 Stroke 2 Normal
PV
(%
)
3D percent vascularity (color Doppler)
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Human imaging with Microultrasound and
Photoacoustics
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Human Imaging
• Ultrasound/Photoacoustic transducer mounted to a 3D stepper motor
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3D Imaging of Human Vasculature
• 3D MIP of the palm • 3D MIP of the forearm
• 3D MIP of the index finger
40mm
20mm
40mm
20mm
• 2D image of the forearm
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Finger Imaging - Ischemia
• Dental floss tied around index finger to induce ischemia
• 2D imaging of finger ischemia – Oxyhemo mode
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Finger Imaging - Ischemia
• 2D imaging of finger ischemia –Spectro mode
Abs.
Figure modified from Murkin and Arango. Near-infrared spectroscopy as an index of brain and tissue Oxygenation. Br J Anaesth 2009; 103 (Suppl. 1): i3–i13
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Imaging of ???
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Doppler imaging of dermatofibroma
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Photoacoustic imaging of dermatofibroma
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Photoacoustic Imaging Co-registered High-resolution High-sensitivity Real-time Oxygen saturation Molecular imaging In Vivo In Situ
LISTEN TO THE LIGHT
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www.visualsonics.com
VisualSonics Insight through in vivo imaging
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